Ascocoryne Sarcoides and Ascocoryne Cylichnium
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Ascocoryne sarcoides and Ascocoryne cylichnium. Descriptions and comparison FINN ROLL-HANSEN AND HELGA ROLL-HANSEN Roll-Hansen, F. & Roll-Hansen, H. 1979. Ascocoryne sarcoides and Ascocoryne cylichnium. Descriptions and comparison. Norw. J. Bot. Vol. 26, pp. 193-206. Oslo. ISSN 0300-1156. Descriptions are given and characters evaluated of apothecia and cultures of Ascocoryne sarcoides (Jacq. ex S. F. Gray) Groves & Wilson and A. cylichnium (L. R. Tul.) Korf. The separation into the two species may be justified by the occurrence of typical ascoconidiain/4. cylichnium, by a relatively sharp boundary between ectal and medullary excipulum in^4. sarcoides, and to some extent by other characters. F. Roll-Hansen & H. Roll-Hansen, Norwegian Forest Research Institute, Forest Pathology, P. O. Box 62, 1432 As, Norway. Ascocoryne species have been isolated by many Former descriptions authors from wood in living spruce trees both in North America and in Europe. In Norway the Ascocoryne sarcoides (Jacq. ex S. F. Ascocoryne species seem to be more common Gray) Groves & Wilson than any other group of fungi in unwounded Groves & Wilson (1967) gave the diagnosis of the stems oiPicea abies. They do not seem to cause genus Ascocoryne and transferred Octospora any rot or discoloration of importance, but they sarcoides Jacq. ex S. F. Gray (Coryne sarcoides may influence the development of other fungi in [Jacq. ex S. F. Gray] Tul.) to that genus. The the stems. conidial state is Coryne dubia Pers. ex S. F. Great confusion exists regarding identification Gray (Pirobasidium sarcoides Hohn.). Descrip of isolates of the Ascocoryne species. In a pre tions of the apothecia with the asci and the liminary paper (Roll-Hansen & Roll-Hansen ascospores have been given by, for example, 1976) the authors reported isolation of Knapp (1924), Dennis (1956), Gremmen (1960), Ascocoryne sarcoides (Jacq. ex S. F. Gray) Christiansen (1962), and Jahn (1967). The apo Groves & Wilson and A. cylichnium (L. R. Tul.) thecia are very similar to those of^4. cylichnium. Korf from stems of Picea abies. Having found al- What Thind & Singh (1969) described as Coryne lantoid conidia in A. sarcoides ascospore- cylichnium (Tul.) Boud. is apparently A. cultures and ovoid conidia in A. cylichnium sarcoides. ascospore-cultures the authors based the Apothecia and conidia-producing stromata in identification of the cultures from the spruce nature. - Knapp (1924) reported that the apo stems on these criteria. But later we found both thecia are 8-13 mm wide. Dennis (1956) stated ovoid and allantoid conidia in varying propor that the apothecia are 2-10 mm across, the tions in some of the cultures. We therefore paraphyses unbranched, about 1 ^.m thick, realized that a closer comparison between apo either enlarged to 2 /xm or abruptly swollen to 4 thecia of A. sarcoides and A. cylichnium, and of fim at the tip, and that the size of the asci is ascospore-cultures from the same ascomata was 90-120 x 8-10 yxm. necessary. The ascospores have been described as el In the present paper we first present a short lipsoid, 1-4-celled, and considerably smaller review of some literature data relevant to dis than in A. cylichnium. According to Knapp tinguishing between the two species. Afterwards (1924) they are 12-18x4-5 ^m; besides the our own material is treated, and a comparison normal ascospores Knapp also found egg-shaped based on literature and our own data is given. conidia in the asci. Dennis (1956) gave the fol- lowing description: 'Ascospores uniseriate or of the apothecia were given, for example, by biseriate, elliptical or inequilateral, at first non- Knapp (1924), Dennis (1956), Gremmen (1960), septate with 2 oil drops, ultimately 1-septate, Christiansen (1962), and Jahn (1967). occasionally with 3 septa, 10—15(—19) x 3-5 p, Apothecia and indication of conidia-produc- sometimes germinating in situ by a terminal ing stromata in nature. - Knapp (1924) reported germ-tube'. Excipulum of 'C. cylichnium' in his that the apothecia are 4-25 mm wide and that the Fig. 151 fits A. sarcoides better than it fits A. ascospores are mostly 25-30 x 6-7 ton, spore cylichnium. minimum being 20 x 5 tim. In the conidial state, Coryne dubia Dennis (1956) gave the following description: (Pirobasidium sarcoides), some millimetres 'Apothecia like those of C. sarcoides but often high, club-like, or often rather irregular, conidial larger, up to 15 mm. across. Flesh of slender stromata are formed. They have the same colour hyaline hyphae, about lti thick, loosely interwo and consistence as the ascomata. The conidia ven in a colourless gelatinous matrix. Excipulum are usually ca. 3.5-6x1 ttm (Hohnel 1902; 40-50 fi thick, of more or less isodiametric, Knapp 1924; Dennis 1956; Christiansen 1962). thin-walled, angular cells, about 8-10 p diame Tulasne & Tulasne (1865) stated that ovate (3.5- ter, forming a compact parenchymatous tissue 6.5 x 3 /im) or sometimes rather globose conidia with an irregular outer surface. Asci cylindric- were also formed on the conidial stromata. clavate, long-stalked, conical-truncate at the Hohnel (1902) found, however, that these 'con apex, the pore blue in Melzer's reagent, 8- idia' were partly pear-shaped 'basidia' producing spored, about 140-200 x 10-12 p. Ascospores the true conidia (4 x 1 pm), partly roundish cel- biseriate fusiform or inequilateral, equally lulae from which the 'basidia' had grown out. pointed at each end, ultimately multiseptate and Cultures on malt agar. - Gremmen (1960) often abstricting spherical secondary spores, ab reported globular 'microconidia' in cultures from out 2 p. across, while still in the ascus; (14)—18— ascospores of A. sarcoides. He thus apparently 30 x 4-6 p. Paraphyses slender, filiform, slightly obtained a type of conidia which is otherwise enlarged towards the apex, embedded in colour supposed to be characteristic of A. cylichnium. less gelatine. On fallen logs and stumps (Fig. Berthet (1964) found rod-shaped, slightly bent 151).' In the figure he drew ectal excipulum and conidia, ca. 5 x 1.5 tim, on the surface in an agar part of medullary excipulum, paraphyses, ascus, culture, but ovoid or subspherical conidia, 2- ascospores, and spherical secondary spores; the 4x2 fim in the agar. Delatour (1976) reported a border between ectal and medullary excipulum similar dimorphism, but he did not mention ('excipulum' and 'flesh') is very sharp, and the whether the roundish conidia were formed in the ascospores are acute at both ends. agar substrate. According to the diagnosis given by Tulasne Etheridge (1957) found no reaction zone on the ascospores are 22-26 x 5-6 tim and rounded gallic acid, except a very intense dark brown at both ends. zone in one case. On tannic acid he found no In most descriptions no conidial state (except reaction zone, or in some isolates light to dark ascoconidia) has been mentioned. But Christ brown zones. iansen (1962) reported finds of apothecia of both Coryne cylichnium (Ascocoryne cylichnium) and Coryne sarcoides (Ascocoryne sarcoides) ac companied by Pirobasidium sarcoides Hohnel (Coryne dubia Pers. ex S. F. Gray). Usually C. Ascocoryne cylichnium (L. R. Tul.) Korf dubia is accepted to be the conidial state of A. Peziza cylichnium was described by Tulasne sarcoides only. (1853) and transferred to Coryne by Boudier in Cultures on malt agar. - Gremmen (1960) 1907. The combination Ascocoryne cylichnium shortly described cultures of A. cylichnium on was validated by Korf (1971). It is interesting malt agar: 'Culture-work (No. 273): Cultures that Karsten (1885) did not consider it a separate obtained from ascospores develop alike as species but named it Coryne sarcoides *urnalis Coryne sarcoides. In older cultures characteris (Nyl.) Karst. (Sarcodea sarcoides *urnalis (Nyl.) tic club-shaped excrescences up to 2.5 mm, Karst. in Not. Sallsk. Faun. Flor. fenn. 11: 232, producing 4 x 1.5 p colourless 1-celled, more or 1870; Ombrophila sarcoides *urnalis (Nyl.) less curved conidia, and reddish coloured micro- Karst. in Myc. Fenn. 1: 87, 1871). Descriptions conidialpustules, were observed.' Gremmen ap- parently happened to get a conidial state cor Phenoloxidase production was tested on gallic responding to C. dubia in his cultures from and tannic acid malt agar (Nobles 1948). Growth ascospores of A. cylichnium. rate of the isolates was measured on malt agar in petri dishes in darkness at the respective temp eratures. Margin of the mycelium (hypha ends) was marked on four diameters after a few days and again 8 to 10 days later. Conidia from Some Norwegian collections. cultures were measured in lactophenol-cotton Descriptions of apothecia and blue under oil immersion (Objective Leitz 90 x, cultures n.a. 1.4). Methods Ascocoryne sarcoides Fresh apothecia from collections of A. sarcoides and A. cylichnium were placed into agar slant Material examined. - Four collections of apo tubes or petri dishes to throw their ascospores thecia, thrown-out ascospores, and ascospore- upwards on sterile agar surfaces, and into moist cultures from them were investigated chambers to throw them on clean, flamed slides. thoroughly: one collection from Malus sp., two The ascospores on the agar surfaces provided from Picea abies, and one from Quercus sp. the pure cultures. The ascospores on the slides Microtome sections were made from one of the were mounted in lactophenol-cottonblue for collections from P. abies. In addition one microscoping. In most cases 50 or 20 ascospores ascospore-culture from apothecia on Alnus sp. were measured. Some of the apothecia were and one from apothecia on Be tula sp. were fixed in 70% FA A, embedded in paraffin wax, investigated thoroughly, but in these two cases microtomed 6 to 12 /xm thick in ribbons, stained material was lacking for further descriptions of in safranin-fastgreen and mounted in canada- the apothecia.